WO2012060783A1 - Utilisation de β-d-glucane pour réduire les effets néfastes d'une dépression de la réponse immunitaire induite par un exercice physique excessif - Google Patents
Utilisation de β-d-glucane pour réduire les effets néfastes d'une dépression de la réponse immunitaire induite par un exercice physique excessif Download PDFInfo
- Publication number
- WO2012060783A1 WO2012060783A1 PCT/SK2011/050016 SK2011050016W WO2012060783A1 WO 2012060783 A1 WO2012060783 A1 WO 2012060783A1 SK 2011050016 W SK2011050016 W SK 2011050016W WO 2012060783 A1 WO2012060783 A1 WO 2012060783A1
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- WO
- WIPO (PCT)
- Prior art keywords
- glucan
- exercise
- physical exercise
- excessive physical
- immune response
- Prior art date
Links
- 230000000694 effects Effects 0.000 title claims abstract description 34
- FYGDTMLNYKFZSV-WFYNLLPOSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,3s,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-WFYNLLPOSA-N 0.000 title claims abstract description 28
- 229920002498 Beta-glucan Polymers 0.000 title claims abstract description 28
- 230000028993 immune response Effects 0.000 title claims abstract description 10
- 230000002411 adverse Effects 0.000 title abstract description 9
- 210000000822 natural killer cell Anatomy 0.000 claims abstract description 29
- 229920001503 Glucan Polymers 0.000 claims abstract description 10
- 230000002538 fungal effect Effects 0.000 claims abstract description 5
- 235000001603 Pleurotus ostreatus Nutrition 0.000 claims abstract description 4
- 240000001462 Pleurotus ostreatus Species 0.000 claims abstract description 4
- 238000011156 evaluation Methods 0.000 claims abstract description 3
- 239000002955 immunomodulating agent Substances 0.000 claims abstract description 3
- 229940121354 immunomodulator Drugs 0.000 claims abstract description 3
- 230000002584 immunomodulator Effects 0.000 claims abstract description 3
- 239000000902 placebo Substances 0.000 description 8
- 229940068196 placebo Drugs 0.000 description 8
- 210000000987 immune system Anatomy 0.000 description 6
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 239000012636 effector Substances 0.000 description 3
- 230000036737 immune function Effects 0.000 description 3
- 230000002101 lytic effect Effects 0.000 description 3
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 2
- 229930003268 Vitamin C Natural products 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000004820 blood count Methods 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 210000000207 lymphocyte subset Anatomy 0.000 description 2
- 230000037081 physical activity Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 235000019154 vitamin C Nutrition 0.000 description 2
- 239000011718 vitamin C Substances 0.000 description 2
- AOFUBOWZWQFQJU-SNOJBQEQSA-N (2r,3s,4s,5r)-2,5-bis(hydroxymethyl)oxolane-2,3,4-triol;(2s,3r,4s,5s,6r)-6-(hydroxymethyl)oxane-2,3,4,5-tetrol Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O.OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@@H]1O AOFUBOWZWQFQJU-SNOJBQEQSA-N 0.000 description 1
- 231100000023 Cell-mediated cytotoxicity Toxicity 0.000 description 1
- 206010057250 Cell-mediated cytotoxicity Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 description 1
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 description 1
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 1
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 description 1
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000005890 cell-mediated cytotoxicity Effects 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000020937 fasting conditions Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 239000000367 immunologic factor Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/20—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
- A23L29/269—Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of microbial origin, e.g. xanthan or dextran
- A23L29/271—Curdlan; beta-1-3 glucan; Polysaccharides produced by agrobacterium or alcaligenes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/716—Glucans
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Definitions
- This invention concerns the use of ⁇ -D-Glucan to reduce the adverse effects that are associated with the immune response (immune system) depression induced by stressful and exhausting physical exercise, particularly in top athletes.
- the ⁇ -D-Glucans (functional polysaccharides) belong to a group of biological response modifiers and are involved in bolstering immune function (Sherwood, E.R., Williams, D.L., and DiLuzio, N.R. 1986, J. Biol. Response Mod. 5: 504-526; Browder, W., Williams, D., Pretus, H. 5 Olivero, G., Enrichens, F., Mao, P., et al. 1990, Ann. Surg. 211 : 605-613; Pretus, H.A., Ensley, H.E., McNamee, R.B., Jones, E.L., Browder, I.W., and Williams, D.L. 1991, J.
- the aim of this invention is to reduce the adverse effects associated with the immune system depression induced by stressful and exhausting physical exercise. Disclosure of Invention
- ⁇ -D-Glucan is used as an immunomodulator to reduce the adverse depression of the immune response following excessive physical exercise.
- ⁇ -D-Glucan is a fungal P(l,3/1,6)-D-Glucan prepared from the oyster mushroom.
- the evaluation the immune response was based on the assessment of NK cell activity immediately prior to excessive physical exercise, within 10 minutes following exercise, and following a relaxation period of 50 to 70 minutes post-exercise.
- NK cell activity after a 50- to 70-minute relaxation period following excessive physical exercise was significantly higher in subjects who had taken ⁇ -D-Glucan compared to those who were given a placebo. Furthermore, this activity was approximately equal to that measured prior excessive physical exercise among subjects who had taken ⁇ -D-Glucan; the mean decrease in NK cell activity (pre-exercise level vs. after the relaxation period) was 3.5% in subjects who had taken ⁇ -D-Glucan and 29% in subjects given the placebo.
- NK cell activity after a 50- to 70-minute relaxation period following excessive physical exercise was virtually unaltered in subjects administered ⁇ -D-Glucan.
- the ⁇ -D-Glucan used in the studies described below was in the form of the commercially prepared Imunoglukan, which is derived from the oyster mushroom.
- Exercise in the form of a gradual bicycle test until exhaustion, was performed by each subject, and the following parameters were measured: blood count and lymphocyte subpopulation analysis from relative and absolute counts (with an emphasis on NK cells, i.e., the NK cell subpopulations CD16 ⁇ , CD8 ⁇ , and CD56 ⁇ and NK cell activity), IgA, IgM, IgG, C3, and C4 levels and Cortisol levels. These parameters were measured prior to exercise, 5 minutes after exercise and after one hour of relaxation following exercise. The athletes were also divided into two therapeutic groups.
- glucan group ingested 100 mg of Imunoglukan and 100 mg of vitamin C in the morning under fasting conditions and prior to exercise
- placebo group ingested a placebo (100 mg of fructose and 100 mg of vitamin C). Both groups adhered to this treatment regimen over the course of 2 months, which was concurrent with their summer training preparation. Two athletes did not complete the study for personal reasons; one was a skier, one was a figure skater, and both were in the glucan group.
- Venous blood was sampled using the Vacutainer blood-draw system in the morning before physical exercise and was analysed with respect to the blood counts and leucocyte and lymphocyte subpopulations (4 ml, K 3 EDTA). Meanwhile, NK cell activity was assessed from heparinised blood samples (7 ml, Na 2 Heparin). The results of this assessment served as the baseline values. A second blood sampling was performed 5 minutes after exercise, and a third sampling was collected after one hour of relaxation following exercise. Both samples were subjected to the same analyses as the baseline sample.
- NK cell activity was conducted using a flow cytometer. Each NK cell activity value was stated in lytic units (LU), which indicated the number of effectors necessary to achieve 20% lysis of 10,000 target cells and was expressed per 10 7 effectors (LU 20 /IO 7 effectors). This NK cell activity value was the only parameter for which a statistically significant difference between the two therapeutic groups was observed (Fig. 1). A difference was also observed in the total NK cell number, but this disparity was not statistically significant.
- LU lytic units
- NK cell activity values expressed in lytic units (LU) for both the glucan and the placebo groups are plotted in Figure.
- NK cell activity decreased below the baseline value, by 28% on average, in the placebo group. Accordingly, the depression in the NK cell number during relaxation after previous physical stress was also accompanied by a functional decrease in NK cell activity. In contrast, we did not observe a decrease in NK cell activity below the baseline value after relaxation in the group that had received glucan.
- NK cell activity after the 50- to 70-minute relaxation period following excessive physical exercise increased by an average of 1.8% in subjects who were administered ⁇ -D-Glucan and decreased by an average of 28% in subjects who were given the placebo. Accordingly, NK cell activity in the group who had taken ⁇ -D-Glucan was virtually unaltered by excessive physical exercise.
- the subjects were assessed in a similar manner as previously described; however, they were administered 180 mg of ⁇ -D-Glucan instead of 100 mg.
- the NK cell activity value after the 50- to 70-minute relaxation period following physical exercise for subjects who received ⁇ -D-Glucan was virtually identical to that the NK cell activity that was measured prior to physical exercise.
- This invention can be used primarily in the field of sports medicine, used primarily by athletes, namely by top sportsmen.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Nutrition Science (AREA)
- Epidemiology (AREA)
- Polymers & Plastics (AREA)
- Immunology (AREA)
- Food Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dispersion Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
L'utilisation de β-D-Lucane comme immunomodulateur se traduit par une diminution des effets contraires d'une dépression de la réponse immunitaire due à un exercice physique excessif chez des individus pratiquant couramment ce type d'exercice. Le β-D-glucane décrit ici est un β(1,3/1,6)-D-glucane fongique préparé à partir de la pleurote et est administré par voie buccale à raison de 200 mg maximum par dose, toujours avant le début d'un exercice physique excessif. L'appréciation de la réponse immunitaire repose sur l'évaluation de l'activité des cellules NK réalisée avant l'exercice physique excessif, dans les 10 minutes suivant l'exercice et entre 50 et 70 minutes après l'exercice.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SKPP50049-2010 | 2010-11-06 | ||
| SK50049-2010A SK500492010A3 (sk) | 2010-11-06 | 2010-11-06 | Use of beta-D-glucane for decreasing undesirable effect of depression of immune response to excessive physical strain |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2012060783A1 true WO2012060783A1 (fr) | 2012-05-10 |
Family
ID=45554778
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/SK2011/050016 WO2012060783A1 (fr) | 2010-11-06 | 2011-11-05 | Utilisation de β-d-glucane pour réduire les effets néfastes d'une dépression de la réponse immunitaire induite par un exercice physique excessif |
Country Status (2)
| Country | Link |
|---|---|
| SK (1) | SK500492010A3 (fr) |
| WO (1) | WO2012060783A1 (fr) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3858362A1 (fr) * | 2020-01-29 | 2021-08-04 | Pleuran, s.r.o. | Bêta-glucane utilisé pour renforcer l'immunité anti-tumorale en rémission |
| WO2021152497A1 (fr) * | 2020-01-29 | 2021-08-05 | Pleuran, S.R.O. | Bêta-glucane servant à la modulation de réponse immunitaire en rémission |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005067977A1 (fr) * | 2004-01-14 | 2005-07-28 | Pleuran, S.R.O. | Procede de preparation d'un hydrogel de glucane fongique presentant une activite antibacterienne et immunostimulatrice, et utilisation correspondante |
| DE202004013521U1 (de) * | 2004-08-30 | 2006-01-05 | Almapharm Christian F.A. Botzenhardt Gmbh + Co | Präparat aus beta-Glucan und Nukleotid zur Verabreichung an Tiere |
| WO2009042230A1 (fr) * | 2007-09-27 | 2009-04-02 | Biopolymer Engineering, Inc. Dba Biothera, Inc | Traitement à base de β-glucane des symptômes d'une infection des voies respiratoires supérieures et du bien-être psychologique |
-
2010
- 2010-11-06 SK SK50049-2010A patent/SK500492010A3/sk not_active Application Discontinuation
-
2011
- 2011-11-05 WO PCT/SK2011/050016 patent/WO2012060783A1/fr active Application Filing
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005067977A1 (fr) * | 2004-01-14 | 2005-07-28 | Pleuran, S.R.O. | Procede de preparation d'un hydrogel de glucane fongique presentant une activite antibacterienne et immunostimulatrice, et utilisation correspondante |
| DE202004013521U1 (de) * | 2004-08-30 | 2006-01-05 | Almapharm Christian F.A. Botzenhardt Gmbh + Co | Präparat aus beta-Glucan und Nukleotid zur Verabreichung an Tiere |
| WO2009042230A1 (fr) * | 2007-09-27 | 2009-04-02 | Biopolymer Engineering, Inc. Dba Biothera, Inc | Traitement à base de β-glucane des symptômes d'une infection des voies respiratoires supérieures et du bien-être psychologique |
Non-Patent Citations (3)
| Title |
|---|
| DAVIS ET AL: "Effects of oat beta-glucan on innate immunity and infection after exercise stress", MEDICINE AND SCIENCE IN SPORTS AND EXERCISE, LIPPINCOTT WILLIAMS & WILKINS, BALTIMORE, MD, US, vol. 36, no. 8, 1 January 2004 (2004-01-01), pages 1321 - 1327, XP008127608, ISSN: 0195-9131 * |
| KATARINA BERGENDIOVA ET AL: "Pleuran (Î-glucan from) supplementation, cellular immune response and respiratory tract infections in athletes", EUROPEAN JOURNAL OF APPLIED PHYSIOLOGY, SPRINGER, BERLIN, DE, vol. 111, no. 9, 20 January 2011 (2011-01-20), pages 2033 - 2040, XP019938793, ISSN: 1439-6327, DOI: 10.1007/S00421-011-1837-Z * |
| TALBOTT S AND TALBOTT J: "Effect of BETA 1,3/1,6 GLUCAN on upper respiratory tract infection symptoms and mood state in marathon athletes", J SPORTS SCI MED, vol. 8, 1 December 2009 (2009-12-01), pages 509 - 519, XP009158119 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3858362A1 (fr) * | 2020-01-29 | 2021-08-04 | Pleuran, s.r.o. | Bêta-glucane utilisé pour renforcer l'immunité anti-tumorale en rémission |
| WO2021152497A1 (fr) * | 2020-01-29 | 2021-08-05 | Pleuran, S.R.O. | Bêta-glucane servant à la modulation de réponse immunitaire en rémission |
Also Published As
| Publication number | Publication date |
|---|---|
| SK500492010A3 (sk) | 2012-06-04 |
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